Microtubule Dynamics and Chromosome Segregation

项目摘要 这个项目的主要目标是了解力的机械贡献。 运动蛋白相关马达和动态微管对染色体的产生 种族隔离。微管和马达都是抗癌的极佳靶点。 但要做出明智的选择来发展治疗学，有必要了解 详细介绍了它们对细胞分裂的贡献。我们已经确定了两个相对未被研究的 与Kinesin相关的马达、Kif25和Kif9，有助于促进适当的关键活动 有丝分裂纺锤体功能。我们将使用高分辨率的活细胞成像，基因编辑， 雷帕霉素依赖的侧写法和体外单分子检测 研究Kif25如何抑制早熟的中心体分离以及它是如何发挥作用的 有丝分裂纺锤体的内向力足以激活纺锤体组装检查点。 我们将使用类似的技术来了解Kif9如何使细胞分裂绕过 主轴组件检查点。这些活动的背后是动态的 作为激动素活性底物的微管。我们已经开发出了一些 使我们能够量化微管动力学的细微变化的分析方法。稳定 微管组装动力学的改变通常是由于丢失或 激动素样调制子或其他肿瘤发生的驱动因素的过度表达。我们将使用 基于细胞的分析和活体成像来确定改变的机制 微管组装速度影响染色体的长期不稳定性。

Project Summary The broad goal of this project is to understand the mechanistic contribution of force generation by kinesin-related motors and dynamic microtubules to chromosome segregation. Both microtubules and motors represent excellent targets for anti-cancer drugs but to make wise choices for developing therapeutics it is necessary to understand their contribution to cell division in detail. We have identified two relatively unstudied kinesin-related motors, Kif25 and Kif9 that contribute key activities to facilitate proper mitotic spindle function. We will use high resolution live cell imaging, gene editing, Rapamycin-dependent knock-sideways approaches and single molecule assays in vitro to investigate how Kif25 suppresses premature centrosome separation and how it exerts inward force on the mitotic spindle sufficient to activate the spindle assembly checkpoint. We will use similar techniques to understand how Kif9 enables dividing cells to bypass the spindle assembly checkpoint. Underlying these activities are the dynamic microtubules that serve as substrates for kinesin activity. We have developed a number of assays that allow us to quantify subtle changes in microtubule dynamics. Stable alterations in microtubule assembly dynamics are often result from loss or overexpression of kinesin-like modulators, or other drivers of tumorigenesis. We will use cell-based assays and live imaging to determine the mechanism by which altered microtubule assembly rates impact long-term chromosome instability.